title: Development of an enzyme-linked immunosorbent assay based on the murine leukemia virus p30 capsid protein.
abstract: Retroviral vectors derived from the murine leukemia virus (MuLV) are widely used as the starting material in the development of vectors for gene therapy and critical in answering questions relating to viral pathogenesis. The p30 capsid (CA) is the major viral core protein and an internal group antigen in MuLV. In this study, an enzyme-linked immunosorbent assay (ELISA) was developed for quantitation of MuLV infectious particles with p30 CA core antigen protein. The ELISA was developed using several goat-polyclonal serum against MuLV p30 generated by the NCI as primary antibody and a rat-monoclonal antibody to CA available from ATCC. The MuLV p30 CA antigen was standardized against recombinant MuLV p30 CA expressed from bacteria. The assay is sensitive, accurate and linear within a defined concentration range of CA. Comparison with different MuLV quantitative methods including reporter gene transfer, reverse transcriptase activity assay, and viral RNA quantitative PCR, showed this ELISA protocol to be highly quantifiable within defined ranges, which can be correlated with infectious viral titer.
authors.affiliation: Department of Pharmacology, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway, NJ 08854 USA. Electronic address: wuta@umdnj.edu.
authors.forename: Dai-Tze
authors.initials: DT
authors.lastname: Wu
authors.forename: Sriram
authors.initials: S
authors.lastname: Aiyer
authors.forename: Rodrigo A
authors.initials: RA
authors.lastname: Villanueva
authors.forename: Monica J
authors.initials: MJ
authors.lastname: Roth
mesh_terms: D000818:Animals; D000914:Antibodies, Viral; D036022:Capsid Proteins / Q000032:analysis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006041:Goats; D009052:Leukemia Virus, Murine / Q000276:immunology / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D019562:Viral Load / Q000379:methods*
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins
keywords: MuLV CA ELISA; Virus normalization; p30 CA